叉头框蛋白A2对肝癌细胞增殖和成瘤的抑制作用

Inhibitory effect of FOXA2 on proliferation and tumorigenesis of hepatocellular carcinoma cells

目的通过研究叉头框蛋白A2(FOXA2)对肝细胞癌细胞体外增殖能力和裸鼠成瘤的影响,探讨FOXA2对肝细胞癌发生发展的影响。方法免疫组化、实时定量PCR检测35对肝细胞癌组织及其配对癌旁组织中FOXA2的表达差异;以293T细胞为对照,利用实时定量PCR检测肝细胞癌细胞系(HepG2、SMMC-7721、SK-Hep1)中FOXA2的表达水平;慢病毒转染HepG2细胞,将实验分为3组:未加病毒组(Mock组)、感染阴性对照慢病毒组(NC组)以及感染FOXA2过表达慢病毒组(FOXA2组);平板克隆实验检测FOXA2对HepG2细胞体外增殖能力的影响;裸鼠成瘤实验检测FOXA2过表达慢病毒感染细胞后对瘤体质量的影响。结果免疫组化、实时定量PCR实验结果表明,癌组织中FOXA2的表达显著低于癌旁组织(均P <0.01);FOXA2在肝癌细胞系(HepG2、SMMC-7721、SK-Hep1)中的表达均显著低于293T细胞(均P <0.0001);FOXA2过表达慢病毒载体转染HepG2细胞后,平板克隆形成实验显示,FOXA2组细胞克隆数较Mock组和NC组明显减少(均P <0.05);裸鼠成瘤实验显示,FOXA2组瘤体质量小于相应空白对照组和阴性对照组(均P <0.01)。结论 FOXA2在肝细胞癌组织和细胞中低表达,具有抑制HepG2细胞体外增殖和裸鼠体内瘤体生长的作用。

Objective To investigate the effects of FOXA2 on the proliferation of hepatocellular carcinoma cells and the tumorigenesis of nude mice, and to explore the effect of FOXA2 on the development of hepatocellular carcinoma. Methods Immunohistochemistiy and real-time quantitative PCR were used to detect the expression of FOXA2 in 35 pairs of hepatocellular carcinoma tissues and their matched paracancerous tissues. 293 T cells were used as controls to detect the expression level of FOX A 2 in hepatocellular carcinoma cell lines(HepG2,SMMC-7721 and SK-Hep1) by real-time quantitative PCR. The lentivirus was transfected into HepG2 cells, and there were 3 groups including no virus group(Mock group), negative control virus group(NC group) and FOXA2-transfected over-expression virus group( FOXA2 group). Plate clone assays were used to detect the effect of FOXA2 on the proliferation of HepG2 cells in vitro and nude mice tumor and formation assays to detect the tumor weight and tumor weight inhibition rate after FOX A2-transfected overexpression of lentivirus-infected cells. Results The results of immunohistochemistry and real-time quantitative PCR showed that the expression of FOXA2 in cancer tissues was significantly lower than that in adjacent tissues(P < 0.01), And the expression of FOXA2 in hepatoma cell lines(HepG2, SMMC-7721, SK-Hepl) was significantly lower than that of 293 T cells(P < 0.0001). After the lentivirus was transfected into HepG2 cells, the number of clones in the FOXA2 group was significantly less than that in the Mock group and the NC group(P < 0.05). The tumor formation of nude mice showed that the tumor weight of FOXA2 group was smaller than that of the corresponding blank control group and negative control group(P < 0.01).Conclusion FOXA2 is lowly expressed in hepatocellular carcinoma tissues and cells, which has the effect of inhibiting the proliferation of HepG2 cells in vitro and the growth of tumors in nude mice in vivo.